OBJECTIVE: We have introduced a method of characterization of invading glioma cells by using molecular analysis of marginal invading tumor cells and molecular profiles of glioma tumor margin. METHODS: Each of tumor core and marginal tissues was obtained in 22 glioma patients. Tumor core cells and marginal cells from each glial tumor were collected by laser capture microdissection or intraoperative microdissection under the operating microscope. Expression of MMP-2, MMP-9, CD44 and RHAMM mRNA by invading glioma cells compared with tumor core was confirmed by realtime-PCR of twenty-four glioma specimens. Clinical data also were reviewed for invasion and recurrence pattern of the gliomas radiologically and invasive rim pattern microscopically. RESULTS: Overall results of the molecular analysis showed that relative overexpression of MMP-2, MMP-9 and RHAMM were noted at the invasive edge of human glioma specimens comparing to the tumor core but CD44 was highly expressed in the tumor core comparing to the margin. High marginal expression of MMP-2 and MMP-9 were noted in poorly ill-defined margin on the pathological finding. High marginal expression of CD44 and MMP-2 were demonstrated in the midline cross group on the radiological review, and that of RHAMM and MMP-2 were showed in the aggressive recurrence group. High expression of MMP-2 seems to be involved in the various invasion-related phenomenons. CONCLUSION: Up-regulation of MMP-2, MMP-9, CD44 and RHAMM was noted in invasive edge of gliomas according to the various clinical situations.
Glioma* ; Humans ; Laser Capture Microdissection ; Microdissection ; Recurrence ; RNA, Messenger ; Up-Regulation

OBJECTIVE: This study was performed to determine the exact pattern of FHIT expression of the cervical carcinoma cell per se by microdissection and to investigate the clinical significance of the FHIT alteration in cervical cancer. METHODS: RT-PCR for FHIT transcript was performed in 18 cervical cancer tissues. Microdissection was performed using laser capture microdissection device and RNA was extracted by RT-nested PCR. PCR products were compared with known aberrant FHIT transcripts. Immunohistochemical analysis was performed to evaluate correlation between the altered expression of FHIT protein and clinical parameters. RESULTS: Six different size of aberrant FHIT transcripts were observed in cervical cancer tissues. Six of 18 (33.3%) cervical cancer sections exhibited full-length normal FHIT transcript only. Aberrant FHIT transcripts with normal one were observed in 9 (50%) and only aberrant transcripts in 3 (16.7%) frozen sections. Five normal cervical tissues expressed only a normal FHIT transcript. The sequences of the 6 different sizes of aberrant FHIT transcripts showed (1) deletion of exons 4-8, (2) deletion of exons 4-7, (3) deletion of exons 5-8, (4) deletion of exons 5-7, (5) deletion of exons 5-7 and insertion of intronic sequences, 153 bp, (6) deletion of exons 5-7 and insertion of intronic sequences, 84 bp. Microdissection of paired cervical tumor and normal stroma showed expression of aberrant FHIT transcripts only in tumor tissues. CONCLUSION: Aberrant FHIT expression was observed frequently in cervical carcinoma and they were observed mainly in cervical cancer cells by microdissection, but not in normal stromal cells. However, absence of FHIT expression did not correlate with clinical prognostic factors in cervical carcinoma.
Exons ; Frozen Sections ; Introns ; Laser Capture Microdissection ; Microdissection* ; Polymerase Chain Reaction ; RNA ; Stromal Cells ; Uterine Cervical Neoplasms*

No abstract available.
Animals ; DNA, Complementary* ; Estrogens ; Laser Capture Microdissection* ; Mice* ; Oligonucleotide Array Sequence Analysis* ; Uterus*

No abstract available.
Animals ; DNA, Complementary* ; Estrogens ; Laser Capture Microdissection* ; Mice* ; Oligonucleotide Array Sequence Analysis* ; Uterus*

No abstract available.
Animals ; Epithelium* ; Gene Expression* ; Laser Capture Microdissection* ; Mice* ; Microarray Analysis* ; Phenobarbital*

In recent years, laser microdissection followed by mass spectrometry (LMD/MS) has been successfully applied to the proteomic studies of formalin-fixed paraffin-embedded (FFPE) renal tissues. This new technique improves the diagnosis of kidney diseases and has a better potential for future clinical application. The review focuses on the use of this methodology for exploring the mechanisms, diagnosis and classification of kidney diseases including renal amyloidosis and membrane proliferative glomerulonephritis.
Formaldehyde ; Humans ; Kidney ; pathology ; Kidney Diseases ; diagnosis ; Laser Capture Microdissection ; Mass Spectrometry ; Proteomics ; Tissue Fixation

Basic research in life science and medicine has dug into single cell level in recent years. Single-cell analysis offers to understand life from diverse perspectives and is used to profile cell heterogeneity to investigate mechanism of diseases. Single cell technologies have also found applications in forensic medicine and clinical reproductive medicine, while the techniques are rapidly evolving and have become more and more sophisticated. In this article, we reviewed various single cell isolation techniques and their pros and cons, including manual cell picking, laser capture microdissection and microfluidics, as well as analysis methods for DNA, RNA and protein in single cell. In addition, we summarized major up-to-date single cell research achievements and their potential applications.
Animals ; Cell Separation ; DNA ; Laser Capture Microdissection ; RNA ; Single-Cell Analysis

The leaner mouse carries a mutation in the gene encoding the alpha1A subunit of P/Q-type calcium channels. Leaner mice exhibit extensive cerebellar granule and Purkinje cell loss that results in cerebellar dysfunction. A previous study suggested that a small population of leaner Purkinje cells undergo apoptosis, however the cell death mode of the rest of degenerating Purkinje cells has not been identified. In order to investigate the mechanisms underlying leaner Purkinje cell death, gene arrays that contain 243 cell death related genes were carried out. To increase the chance of detecting Purkinje cell specific genes, laser capture microdissection was employed to obtain Purkinje cell enriched samples. The gene array analysis revealed several potential genes that are involved in autophagic cell death pathway including cathepsin D, a key lysosomal protease that triggers autophagic degradation. Further analysis on LC3, which is a hallmark for autophagic cell death showed that leaner Purkinje cells are degenerating via autophagic process. The present study provides evidence that calcium channel defects trigger different modes of neurodegeneration in the cerebellum.
Animals ; Apoptosis ; Autophagy ; Calcium Channels ; Cathepsin D ; Cell Death ; Cerebellar Diseases ; Cerebellum ; Laser Capture Microdissection ; Mice ; Purkinje Cells

The leaner mouse carries a mutation in the gene encoding the alpha1A subunit of P/Q-type calcium channels. Leaner mice exhibit extensive cerebellar granule and Purkinje cell loss that results in cerebellar dysfunction. A previous study suggested that a small population of leaner Purkinje cells undergo apoptosis, however the cell death mode of the rest of degenerating Purkinje cells has not been identified. In order to investigate the mechanisms underlying leaner Purkinje cell death, gene arrays that contain 243 cell death related genes were carried out. To increase the chance of detecting Purkinje cell specific genes, laser capture microdissection was employed to obtain Purkinje cell enriched samples. The gene array analysis revealed several potential genes that are involved in autophagic cell death pathway including cathepsin D, a key lysosomal protease that triggers autophagic degradation. Further analysis on LC3, which is a hallmark for autophagic cell death showed that leaner Purkinje cells are degenerating via autophagic process. The present study provides evidence that calcium channel defects trigger different modes of neurodegeneration in the cerebellum.
Animals ; Apoptosis ; Autophagy ; Calcium Channels ; Cathepsin D ; Cell Death ; Cerebellar Diseases ; Cerebellum ; Laser Capture Microdissection ; Mice ; Purkinje Cells

Amyloidosis is a heterogeneous group of diseases caused by deposition of misfolded proteins, which usually leads to organ dysfunction. Accurate typing of amyloid deposits is of paramount importance because organ involvements and disease prognosis differ widely among different subtypes, and its treatments are type specific. Correct identification of amyloidogenic protein is crucial to proper treatment. Traditional antibody-based diagnostic methods such as immunohistochemistry and immunofluorescence are helpful in amyloid typing, but limitations of those approaches including antibody availability and serum protein contamination impair sensitivity and specificity of diagnosis. Sometimes misdiagnosis can lead to catastrophic therapeutic outcome. Genetic testing is important to confirm the diagnosis of hereditary amyloidosis. Nowadays proteomic analysis has been used as an advanced strategy for amyloid typing and the gold-standard today is laser microdissection followed by mass spectrometry (LMD/MS), which can identify causal protein without additional clinical information. Furthermore, LMD/MS is performed on formalin-fixed paraffin-embedded (FFPE) specimens, thus large scale retrospective studies based on archival material can be conducted. In recent studies, LMD/MS has been proven superior to traditional methods without the drawbacks mentioned above. This proteomic approach provides guarantee of appropriate clinical management and probability of new insights into the mechanism of amyloidosis.In this article the new advances of studies on subtyping of systemic amyloidosis are reviewed.
Amyloidosis ; classification ; pathology ; Humans ; Immunoglobulin Light-chain Amyloidosis ; Laser Capture Microdissection ; Mass Spectrometry ; Plasma Cells ; Proteomics ; methods